条斑紫菜蛋白酶解产物中α-葡萄糖苷酶抑制剂纯化及特性  

Purification and Characterization of α-Glucosidase Inhibitor from Proteolytic Product of Porphyra Yezoensis

作　　者：周锋[1] 刘冬冬[1] 周楠迪[1] 田亚平[1] 

机构地区：[1]江南大学工业生物技术教育部重点实验室,无锡214122

出　　处：《天然产物研究与开发》2014年第9期1463-1468,共6页Natural Product Research and Development

基　　金：江苏省农业支撑项目(BE2013358)

摘　　要：以α-葡萄糖苷酶抑制活性为指标,优选出1398中性蛋白酶、碱性蛋白酶和氨肽酶在一定条件下复配酶解条斑紫菜蛋白制备α-葡萄糖苷酶抑制剂。酶解液加入乙醇至终浓度为60%以沉淀去除多糖,上清液为α-葡萄糖苷酶抑制剂粗品。该粗品利用SP Sepharose High Performance阳离子交换层析、Sephadex G-10凝胶层析和Mono Q阴离子交换层析进行分离纯化,获得一种肽类α-葡萄糖苷酶抑制剂(LGI)。LGI经反向高效液相层析测定纯度为62.4%,基本特性分析显示其具较好的温度和pH稳定性,对α-葡萄糖苷酶半抑制浓度IC50值为97.36μg/mL,属于一种非竞争性抑制剂。In this study, combined proteolyzation of Porphyra yezoensis by 1398 neutral protease, alkaline protease and aminopeptidase was optimized using the a-glueosidase inhibitor activity as index. The a-glueosidase inhibitor from hy- drolysate was initially extracted by adding 60% ethanol to precipitate and remove polysaccharides. The supematant was further purified through SP Sepharose High Performance cation-exchange chromatography, Sephadex G-10 gel filtration chromatography and Mono Q anion exchange chromatography. The purity of the obtained peptide-type α-glucosidase in- hibitor （LGI） was up to 62.4% ,according to the result of reverse-phase high performance liquid chromatography （ RP- HPLC）. The basic characteristics of LGI indicated that it had high thermo-stability and pH stability. The IC50value of LGI was 97.36 μg/mL against a-glucosidase, and it belonged to non-competitive inhibitors.

关 键 词：条斑紫菜 蛋白酶水解 Α-葡萄糖苷酶抑制剂 纯化 基本特性 

分 类 号：Q946.1[生物学—植物学] TS254.9[轻工技术与工程—水产品加工及贮藏工程]